This proposal contains experiments designed to study the interaction of Neisseria gonorrhoeae with the contents of phagolysosomes isolated from human neutrophils and monocytes/macrophages. The objective of the proposed experimental plan is to study, on a biochemical and bacteriological level, the processes involved in the bactericidal activity expressed by human leukocytes toward N. gonorrhoeae and to define the damage suffered by N. gonorrhoeae. It is hoped that these studies will lead to a better understanding of acute gonorrhea and thus to a better understanding of how to treat the disease and prevent its debilitating and sometimes fatal complications. Initial experiments involve isolating phagolysosomes from human leukocytes by allowing purified neutrophils or monocytes to phagocytize buoyant, non-digestible particles (latex, polyvinyl toluene or paraffin oil emulsions), homogenizing the leukocytes and separating the subcellular components through sucrose gradients. Various biologically active materials will be used to induce phagolysosome formation and the bactericidal capacity of phagolysosomal contents extracted by several different methods will be compared. Phagolysosomal extracts will be quantitatively and qualitatively measured for content of proteins, both enzymatic and non-enzymatic, thought to be involved in either oxidative or non-oxidative bactericidal activity, including myeloperoxidase, lysozyme, lactoferrin, collagenase, elastase, chymotrypsin-like cationic proteins, proteases, glycosidases and phospholipases. To see if growth conditions affect acquisition of virulence factors, N. gonorrhoeae will be grown both in vitro on bacteriologic media and in vivo in subcutaneous chambers implanted in guinea pig backs or in allantoic fluid of chicken embryos. Structural, physiological and biochemical consequences of the degradation or killing of N. gonorrhoeae by phagolysosomal extracts will be studied, and the enzymatic and biochemical mechanisms of the observed degradation and killing will be elucidated.